Extracellular cathepsin S and intracellular caspase 1 activation are surrogate biomarkers of particulate-induced lysosomal disruption in macrophages.
Particulate matter has been shown to stimulate the innate immune system and induce acute inflammation. Therefore, while nanotechnology has the potential to provide therapeutic formulations with improved efficacy, there are concerns such pharmaceutical preparations could induce unwanted inflammatory side effects. Accordingly, we aim to examine the utility of using the proteolytic activity signatures of cysteine proteases, caspase 1 and cathepsin S (CTSS), as biomarkers to assess particulate-induced inflammation. Primary peritoneal macrophages and bone marrow-derived macrophages from C57BL/6 mice and ctss(-/-) mice were exposed to micro- and nanoparticulates and also the lysosomotropic agent, L-leucyl-L-leucine methyl ester (LLOME). ELISA and immunoblot analyses were used to measure the IL-1β response in cells, generated by lysosomal rupture. Affinity-binding probes (ABPs), which irreversibly bind to the active site thiol of cysteine proteases, were then used to detect active caspase 1 and CTSS following lysosomal rupture. Reporter substrates were also used to quantify the proteolytic activity of these enzymes, as measured by substrate turnover. We demonstrate that exposure to silica, alum and polystyrene particulates induces IL-1β release from macrophages, through lysosomal destabilization. IL-1β secretion positively correlated with an increase in the proteolytic activity signatures of intracellular caspase 1 and extracellular CTSS, which were detected using ABPs and reporter substrates. Interestingly IL-1β release was significantly reduced in primary macrophages from ctss(-/-) mice. This study supports the emerging significance of CTSS as a regulator of the innate immune response, highlighting its role in regulating IL-1β release. Crucially, the results demonstrate the utility of intracellular caspase 1 and extracellular CTSS proteolytic activities as surrogate biomarkers of lysosomal rupture and acute inflammation. In the future, activity-based detection of these enzymes may prove useful for the real-time assessment of particle-induced inflammation and toxicity assessment during the development of nanotherapeutics.


Factors Involved in the Emergence and Persistence of Food-Borne Diseases †.
In recent years, a number of bacteria, viruses, and parasites have emerged as food-borne pathogens and resulted in numerous food-borne disease outbreaks. These outbreaks have had a major impact in terms of loss of human lives and economic costs. Genetic changes in microorganisms resulting in increased virulence, changes in social attitudes and eating habits, changes in food production and distribution systems, an increase in the number of immunocompromised individuals, and improved pathogen-detection methods are some of the factors that have contributed to the emergence/recognition and persistence of food-borne pathogens. The causes leading to the emergence of new food-borne pathogens or the reemergence of pathogens involve the interaction of several factors. This review discusses in detail factors involved in the emergence/recognition and persistence of several bacterial, parasitic, viral, and virus-like agents associated with food-borne diseases of public-health significance.


Effect of PCI on quality of life in patients with stable coronary disease.
It has not been clearly established whether percutaneous coronary intervention (PCI) can provide an incremental benefit in quality of life over that provided by optimal medical therapy among patients with chronic coronary artery disease. We randomly assigned 2287 patients with stable coronary disease to PCI plus optimal medical therapy or to optimal medical therapy alone. We assessed angina-specific health status (with the use of the Seattle Angina Questionnaire) and overall physical and mental function (with the use of the RAND 36-item health survey [RAND-36]). At baseline, 22% of the patients were free of angina. At 3 months, 53% of the patients in the PCI group and 42% in the medical-therapy group were angina-free (P<0.001). Baseline mean (+/-SD) Seattle Angina Questionnaire scores (which range from 0 to 100, with higher scores indicating better health status) were 66+/-25 for physical limitations, 54+/-32 for angina stability, 69+/-26 for angina frequency, 87+/-16 for treatment satisfaction, and 51+/-25 for quality of life. By 3 months, these scores had increased in the PCI group, as compared with the medical-therapy group, to 76+/-24 versus 72+/-23 for physical limitation (P=0.004), 77+/-28 versus 73+/-27 for angina stability (P=0.002), 85+/-22 versus 80+/-23 for angina frequency (P<0.001), 92+/-12 versus 90+/-14 for treatment satisfaction (P<0.001), and 73+/-22 versus 68+/-23 for quality of life (P<0.001). In general, patients had an incremental benefit from PCI for 6 to 24 months; patients with more severe angina had a greater benefit from PCI. Similar incremental benefits from PCI were seen in some but not all RAND-36 domains. By 36 months, there was no significant difference in health status between the treatment groups. Among patients with stable angina, both those treated with PCI and those treated with optimal medical therapy alone had marked improvements in health status during follow-up. The PCI group had small, but significant, incremental benefits that disappeared by 36 months. (ClinicalTrials.gov number, NCT00007657.)


Treatment of pediatric atlantoaxial instability with traditional and modified Goel-Harms fusion constructs.
There are several treatment options for rigid fixation at C1-C2 including Brooks and Gallie type wired fusions and C1-2 transarticular screws. The use of a Goel-Harms type fusion, a construct with C1 lateral mass screws and C2 pedicle screws, has not been extensively described in pediatric patients. Here, we describe its relatively safe and effective use for treating pediatric patients by retrospective chart review of patients treated by the senior author for atlantoaxial instability with a Goel-Harms-type constructs during a 3-year period (2005-2007). Six patients were treated using Goel-Harms-type constructs. Five patients were treated utilizing a construct containing C1 lateral mass screws and C2 pedicle screws; one patient was treated using construct containing C1 lateral mass screws and C2 trans-laminar screws. The patients ranged in age from 7 to 17 years old (mean 12.7). All patients had findings of an os odontoideum on CT scans and three of the six patients had T2 hyperintensity on MRI. Three of the six patients presented with transient neurologic deficits: quadraplegia in two patients and paresthesias in two patients. In each patient C1 lateral mass and C2 screws were placed and the subluxation was reduced to attain an anatomical alignment. No bone grafts were harvested from the iliac crest or rib. Local morsalized bone and sub-occipital skull graft was used. All patients tolerated the procedure well and were discharged home on post-operative day 3-4. The patients wore a hard cervical collar and no halo-vests were needed. All patients had solid fusion constructs and normal alignment on post-operative imaging studies performed on average 14 months post-operatively (range: 7-29). The results demonstrated that Goel-Harms fusions are a relatively safe and effective method of treating pediatric patients with atlantoaxial instability and are not dependent on vertebral anatomy or an intact ring of C1. Follow-up visits and studies in this limited series of patients demonstrated solid fusion constructs and anatomical alignment in all patients treated.


A retrospective international study on primary extranodal marginal zone lymphoma of the lung (BALT lymphoma) on behalf of International Extranodal Lymphoma Study Group (IELSG).
Primary lymphoma of the lung is a rare entity. Clinical features, optimal treatment, role of surgery and outcomes are not well defined, and the follow-up is variable in published data. Clinical data of 205 patients who were confirmed to have bronchus mucosa-associated lymphoid tissue lymphoma from December 1986 to December 2011 in 17 different centres worldwide were evaluated. Fifty-five per cent of the patients were female. The median age at diagnosis was 62 (range 28-88) years. Only 9% had a history of exposure to toxic substances, while about 45% of the patients had a history of smoking. Ten per cent of the patients had autoimmune disease at presentation, and 19% patients had a reported preexisting lung disease. Treatment modalities included surgery alone in 63 patients (30%), radiotherapy in 3 (2%), antibiotics in 1 (1%) and systemic treatment in 128 (62%). Patients receiving a local approach, mainly surgical resection, experienced significantly improved progression-free survival (p = 0.003) versus those receiving a systemic treatment. There were no other significant differences among treatment modalities. The survival data confirm the indolent nature of the disease. Local therapy (surgery or radiotherapy) results in long-term disease-free survival for patients with localized disease. Systemic treatment, including alkylating-containing regimens, can be reserved to patients in relapse after incomplete surgical excision or for patients with advanced disease. Copyright © 2015 John Wiley & Sons, Ltd.


The major bioactive components of seaweeds and their mosquitocidal potential.
Seaweeds are one of the most widely studied natural resources for their biological activities. Novel seaweed compounds with unique chemical structures have been reported for their pharmacological properties. The urge to search for novel insecticidal compound with a new mode of action for development of botanical insecticides supports the relevant scientific research on discovering the bioactive compounds in seaweeds. The mosquitocidal potential of seaweed extracts and their isolated compounds are documented in this review paper, along with the discussion on bioactivities of the major components of seaweeds such as polysaccharides, phenolics, proteins, terpenes, lipids, and halogenated compounds. The effects of seaweed extracts and compounds toward different life stages of mosquito (egg, larva, pupa, and adult), its growth, development, and reproduction are elaborated. The structure-activity relationships of mosquitocidal compounds are discussed to extrapolate the possible chemical characteristics of seaweed compounds responsible for insecticidal properties. Furthermore, the possible target sites and mode of actions of the mosquitocidal seaweed compounds are included in this paper. The potential synergistic effects between seaweeds and commercial insecticides as well as the toxic effects of seaweed extracts and compounds toward other insects and non-target organisms in the same habitat are also described. On top of that, various factors that influence the mosquitocidal potential of seaweeds, such as abiotic and biotic variables, sample preparation, test procedures, and considerations for a precise experimental design are discussed. The potential of active seaweed extracts and compounds in the development of effective bioinsecticide are also discussed.


Light-emitting diode spherical packages: an equation for the light transmission efficiency.
Virtually all light-emitting diodes (LEDs) are encapsulated with a transparent epoxy or silicone gel. Here we analyze the optical efficiency of spherical encapsulants. We develop a quasi-radiometric equation for the light transmission efficiency, which incorporates some ideas of Monte Carlo ray tracing into the context of radiometry. The approach includes the extended source nature of the LED chip and the chip radiance distribution. The equation is an explicit function of the size and the refractive index of the package, and also of several chip parameters such as shape, size, radiance, and location inside the package. To illustrate the use of this equation, we analyze several packaging configurations of practical interest, for example, a hemispherical dome with multiple chips, a flat encapsulation as a special case of the spherical package, and approximate calculations of an encapsulant with a photonic crystal LED or a photonic quasi-crystal LED. These calculations are compared with Monte Carlo ray tracing, giving almost identical values.


Transgender Youth Experiences with Implantable GnRH Agonists for Puberty Suppression.
This descriptive study reports caregiver experiences with GnRH agonist implants among a cohort of youth followed in a pediatric hospital-based gender clinic. We administered a survey to 36 of 55 eligible caregivers ascertaining demographics and satisfaction, with a medical record review of any surgical complications. The overwhelming majority (97.1%) reported satisfaction with the procedure and would undergo the implant procedure again (94.4%). The most frequent challenges noted were about affordability (39.8%) and insurance denials (39.8%). Implantable GnRH agonist can be used successfully in pediatric patients with gender dysphoria. Future policy should seek to address concerns regarding insurance approval and reimbursement.


Heat shock protein 70 is acute phase reactant: response elicited by tumor treatment with photodynamic therapy.
Oxidative stress in photodynamic therapy (PDT)-treated tumor cells is known to instigate a strong upregulation of the expression of heat shock proteins. However, the treatment of mouse Lewis lung carcinoma (LLC) cells with Photofrin™ PDT resulted in the upregulation of heat shock protein 70 (Hsp70) gene not only in these cells but also in co-incubated untreated Hepa 1-6 cells. To investigate whether this phenomenon extends in vivo, LLC tumors growing in C57BL/6 mice were treated with Photofrin™ PDT. The tumors and the livers from the mice were collected at 4, 8, or 24 h after therapy for quantitative reverse transcriptase polymerase chain reaction-based analysis of Hsp70 gene expression. Increased Hsp70 gene expression was detected in both the tumor and liver tissues and was most pronounced at 4 h after PDT. This effect was inhibited by treatment of host mice with glucocorticoid synthesis inhibitor metyrapone. Hsp70 protein levels in the livers of mice bearing PDT-treated tumors gradually decreased after therapy while serum levels increased at 4 h after therapy and then continually decreased. The exposure of in vitro PDT-treated LLC cells to Hsp70 and subsequent flow cytometry analysis revealed binding of this protein to cells that was dependent on PDT dose and more pronounced with dying than viable cells. Thus, following the induction of tumor injury by PDT, Hsp70 can be produced in the liver and spleen as acute phase reactant and released into circulation, from where it can be rapidly sequestered to damaged tumor tissue to facilitate the disposal of dying cells.


Activation of macrophages in vivo and in vitro. Correlation between hydrogen peroxide release and killing of Trypanosoma cruzi.
As reported previously, mouse peritoneal macrophages could be activated to kill intracellular trypomastigotes of Trypanosoma cruzi, the agent of Chagas' disease, in either of two ways: by immunizing and boosting the mice (3), or by culturing resident or inflammatory macrophages in spleen cell factor(s) (SCF) in vitro (2). Macrophages activated in vivo became less trypanocidal with time in culture, and cells activated in vitro lost trypanocidal capacity when CSF was removed (2). In the present study, the ability of macrophages to release H2O2 in response to phorbol myristate acetate (PMA) could be induced in vivo and in vitro, and reversed in vitro, in a manner correlating closely with changes in trypanocidal activity. Macrophages could be activated in vitro with SCF in a time-dependent and dose-dependent fashion, so that they released as much H2O2 as macrophages activated in vivo. The sensitivity of epimastigotes and trypomastigotes to enzymatically generated H2O2 suggested that the generation of H2O2 by activated macrophages could be plausible explanation for their trypanocidal activity. Of the biochemical correlates of macrophage activation reported to date, increased ability to release H2O2 seems most closely allied to enhanced capacity to kill an intracellular pathogen.


Management of an odontoid synchondrosis fracture causing chronic translational anterior atlanto-axial subluxation in a child with autism: case report.
The authors report an unusual case of an odontoid synchondrosis fracture causing chronic translational anterior atlanto-axial subluxation and present a discussion of the unique management of this case. Traumatic translational anterior atlanto-axial subluxation is a rare manifestation within pediatrics. Patients with preexisting abnormalities in ligamentous or bony structures may present with unusual symptomatology, which could result in delay of treatment. A 6-year-old male patient with autism who presented with acute respiratory arrest was noted to have an odontoid synchondrosis fracture and severe anterior translational atlanto-axial subluxation. Initial attempts at reduction with halo traction were tried for first-line treatment. However, because of concern regarding possible inadvertent worsening of the impingement, the presence of comorbid macrocephaly, and possible instability with only C1-2 fusion, a posterior C1 laminectomy was performed. Further release of the C1-2 complex and odontoid peg from extensive fibrous tissue allowed for complete reduction. Acute injuries of the C1-2 complex may not present as expected, and the presence of pain is not a reliable symptom. Halo traction is an appropriate initial treatment, but some patients may require surgical realignment and stabilization.


Paneth cell marker expression in intestinal villi and colon crypts characterizes dietary induced risk for mouse sporadic intestinal cancer.
Nutritional and genetic risk factors for intestinal tumors are additive on mouse tumor phenotype, establishing that diet and genetic factors impact risk by distinct combinatorial mechanisms. In a mouse model of dietary-induced sporadic small and large intestinal cancer in WT mice in which tumor etiology, lag, incidence, and frequency reflect >90% of intestinal cancer in Western societies, dietary-induced risk altered gene expression profiles predominantly in villus cells of the histologically normal mucosa, in contrast to targeting of crypt cells by inheritance of an Apc(1638N) allele or homozygous inactivation of p21(Waf1/cip1), and profiles induced by each risk factor were distinct at the gene or functional group level. The dietary-induced changes in villus cells encompassed ectopic expression of Paneth cell markers (a lineage normally confined to the bottom of small intestinal crypts), elevated expression of the Wnt receptor Fzd5 and of EphB2 (genes necessary for Paneth cell differentiation and localization to the crypt bottom), and increased Wnt signaling in villus cells. Ectopic elevation of these markers was also present in the colon crypts, which are also sites of sporadic tumors in the nutritional model. Elevating dietary vitamin D(3) and calcium, which prevents tumor development, abrogated these changes in the villus and colon cells. Thus, common intestinal cancer driven by diet involves mechanisms of tumor development distinct from those mechanisms that cause tumors induced by the rare inheritance of a mutant adenomatous polyposis coli (Apc) allele. This is fundamental for understanding how common sporadic tumors arise and in evaluating relative risk in the population.


Evidence of corneal lymphangiogenesis in dry eye disease: a potential link to adaptive immunity?
To determine the effect of desiccating stress on corneal angiogenic responses in dry eye disease (DED) using a murine model. Dry eye was induced in murine eyes using high-flow desiccated air. Corneas were double stained with CD31 (panendothelial marker) and LYVE-1 (lymphatic endothelial marker). Real-time polymerase chain reaction was performed to quantify expression of vascular endothelial growth factors (VEGF-A, VEGF-C, and VEGF-D) and their receptors (VEGFR-2 and VEGFR-3) in the cornea on days 6, 10, and 14. Enumeration of CD11b(+)/LYVE-1(+) monocytic cells was performed in corneas with DED on day 14. Flow cytometric evaluation of the draining lymph nodes in normal mice and mice with DED was performed to determine whether DED is associated with homing of mature (major histocompatibility complex class II(hi)) antigen-presenting cells to the lymphoid compartment. Lymphatic vessels unaccompanied by blood vessels were seen growing toward the center of corneas with DED. Significant increases in lymphatic area (P < .001) and lymphatic caliber (P < .02) were seen on day 14 of disease. Lymphangiogenic-specific VEGF-D and VEGFR-3 levels increased earliest on day 6 followed by increased VEGF-C, VEGF-A, and VEGFR-2 levels. Increased recruitment of CD11b(+)/LYVE-1(+) monocytic cells to the cornea and homing of mature CD11b(+) antigen-presenting cells to the draining lymph nodes were also associated with DED. Low-grade inflammation associated with DED is an inducer of lymphangiogenesis without accompanying hemangiogenesis.


A graduate oral health therapist program to support dental service delivery and oral health promotion in Aboriginal communities in New South Wales, Australia.
The early closure of the Voluntary Dental Graduate Year Program and the Oral Health Therapy Graduate Year Program by the Australian Government adversely impacted New South Wales (NSW) Aboriginal Community Controlled Health Services (ACCHSs). This led to the co-design of a small-scale oral health therapy graduate year program for ACCHSs known as the Dalang Project, which enabled oral health therapists to engage with local Aboriginal communities and implement culturally competent, practical and evidence-based oral health promotion activities. This article provides an overview of the Dalang Project and its evaluation. All graduates of the Dalang Project were invited via email and social media to complete an online survey. The survey included questions about their year in the Dalang Project, why they applied, what they liked and disliked about the project, where they planned to work post-placement, and examples of the most significant changes they observed in the communities where they were placed. Host sites were also surveyed and data were collated on clinical services performed as well as oral health promotion activity. Prior to commencing the Dalang Project only 4 of the 15 respondents came from rural or regional areas. Nine of the 15 respondents were considering working in a regional, rural or remote area prior to applying for the Dalang Project. Twelve respondents were working at the time of the survey and half were working in regional, rural or remote locations in NSW and one in the Northern Territory. All reported that they would be more likely to work in an ACCHS as a result of being a part of the Dalang Project. The majority of respondents said they would recommend the program to future graduates. A total of 63 schools, 21 preschools and 15 community health services received regular dental health education through the Dalang Project. A total of 3250 toothbrushes and fluoride toothpastes were distributed to children and families through the Dalang Project. A key part of the program was the installation of refrigerated and filtered water fountains in schools where there was no free filtered or refrigerated water supply. The inclusion of this component in the program was part of the co-design process and links the program to the wider population health strategies in NSW to help prevent childhood obesity. The Dalang Project is an example of a successful co-designed project that has positively impacted oral health service delivery for Aboriginal children and has provided a valuable experience for new graduate oral health therapists working in ACCHSs. Overall, the Dalang Project was found to be a positive professional experience for the oral health therapists with many remaining in rural, remote and regional locations after completing the program.


The methylated N-terminal tail of RCC1 is required for stabilisation of its interaction with chromatin by Ran in live cells.
Regulator of chromosome condensation 1 (RCC1) is the guanine nucleotide exchange factor for Ran GTPase. Localised generation of Ran-GTP by RCC1 on chromatin is critical for nucleocytoplasmic transport, mitotic spindle assembly and nuclear envelope formation. Both the N-terminal tail of RCC1 and its association with Ran are important for its interaction with chromatin in cells. In vitro, the association of Ran with RCC1 induces a conformational change in the N-terminal tail that promotes its interaction with DNA. We have investigated the mechanism of the dynamic interaction of the alpha isoform of human RCC1 (RCC1alpha) with chromatin in live cells using fluorescence recovery after photobleaching (FRAP) of green fluorescent protein (GFP) fusions. We show that the N-terminal tail stabilises the interaction of RCC1alpha with chromatin and this function can be partially replaced by another lysine-rich nuclear localisation signal. Removal of the tail prevents the interaction of RCC1alpha with chromatin from being stabilised by RanT24N, a mutant that binds stably to RCC1alpha. The interaction of RCC1alpha with chromatin is destabilised by mutation of lysine 4 (K4Q), which abolishes alpha-N-terminal methylation, and this interaction is no longer stabilised by RanT24N. However, alpha-N-terminal methylation of RCC1alpha is not regulated by the binding of RanT24N. Conversely, the association of Ran with precipitated RCC1alpha does not require the N-terminal tail of RCC1alpha or its methylation. The mobility of RCC1alpha on chromatin is increased by mutation of aspartate 182 (D182A), which inhibits guanine-nucleotide exchange activity, but RCC1alphaD182A can still bind nucleotide-free Ran and its interaction with chromatin is stabilised by RanT24N. These results show that the stabilisation of the dynamic interaction of RCC1alpha with chromatin by Ran in live cells requires the N-terminal tail of RCC1alpha. alpha-N-methylation is not regulated by formation of the binary complex with Ran, but it promotes chromatin binding through the tail. This work supports a model in which the association of RCC1alpha with chromatin is promoted by a conformational change in the alpha-N-terminal methylated tail that is induced allosterically in the binary complex with Ran.


In situ control of polymer helicity with a non-covalently bound photoresponsive molecular motor dopant.
The transfer of chirality from a molecular motor to a dynamic helical polymer via ionic interactions was investigated. A dopant with photoswitchable chirality was able to induce a preferred helicity in a poly(phenylacetylene) polymer and the helicity is inverted upon irradiation. The findings described herein will advance the development of functional and responsive polymeric systems.


Glucocorticoid-induced bone loss in dermatologic patients: an update.
To raise awareness of the new treatment options and current recommendations among dermatologists treating patients with systemic corticosteroids. MEDLINE peer-reviewed publications. English language and clinical pertinence to corticosteroid-induced osteoporosis. Pertinent information on pathophysiologic, epidemiologic, clinical trial, cost-effectiveness, and treatment option data regarding corticosteroid-induced osteoporosis. Comprehensive summary of published data on the pathophysiologic, epidemiologic, clinical, and treatment data and current practice guidelines regarding corticosteroid-induced osteoporosis; creation of an algorithmic management approach for patients treated with long-term oral corticosteroids. Glucocorticoid-induced bone loss is the most predictable and debilitating complication of prolonged administration of systemic corticosteroids. Every dermatologist prescribing systemic corticosteroids must be aware of corticosteroid-induced osteoporosis and ensure that every patient is receiving general measures to prevent it. Despite efficacious preventive and therapeutic options, actual implementation of these strategies remains unacceptably low. Based on currently available evidence, the first choice for prevention and treatment of glucocorticoid-induced osteoporosis should be a potent oral bisphosphonate such as alendronate (70 mg/wk) or risedronate sodium (35 mg/wk). For patients with severe osteoporosis or patients with active osteoporotic fractures, the anabolic agent teriparatide (recombinant fragmented parathyroid hormone) should be considered as a first-line option for up to 2 years.


Invasive Macrophytes Control the Spatial and Temporal Patterns of Temperature and Dissolved Oxygen in a Shallow Lake: A Proposed Feedback Mechanism of Macrophyte Loss.
Submerged macrophytes can have a profound effect on shallow lake ecosystems through their ability to modify the thermal structure and dissolved oxygen levels within the lake. Invasive macrophytes, in particular, can grow rapidly and induce thermal gradients in lakes that may substantially change the ecosystem structure and challenge the survival of aquatic organisms. We performed fine-scale measurements and 3D numerical modeling at high spatiotemporal resolution to assess the effect of the seasonal growth of Potamogeton crispus L. on the spatial and temporal dynamics of temperature and dissolved oxygen in a shallow urban lake (Lake Monger, Perth, WA, Australia). Daytime stratification developed during the growing season and was clearly observed throughout the macrophyte bed. At all times measured, stratification was stronger at the center of the macrophyte bed compared to the bed edges. By fitting a logistic growth curve to changes in plant height over time (r2 = 0.98), and comparing this curve to temperature data at the center of the macrophyte bed, we found that stratification began once the macrophytes occupied at least 50% of the water depth. This conclusion was strongly supported by a 3D hydrodynamic model fitted to weekly temperature profiles measured at four time periods throughout the growing season (r2 > 0.78 at all times). As the macrophyte height increased and stratification developed, dissolved oxygen concentration profiles changed from vertically homogeneous oxic conditions during both the day and night to expression of night-time anoxic conditions close to the sediments. Spatially interpolated maps of dissolved oxygen and 3D numerical modeling results indicated that the plants also reduced horizontal exchange with surrounding unvegetated areas, preventing flushing of low dissolved oxygen water out of the center of the bed. Simultaneously, aerial imagery showed central dieback occurring toward the end of the growing season. Thus, we hypothesized that stratification-induced anoxia can lead to accelerated P. crispus dieback in this region, causing formation of a ring-shaped pattern in spatial macrophyte distribution. Overall, our study demonstrates that submerged macrophytes can alter the thermal characteristics and oxygen levels within shallow lakes and thus create challenging conditions for maximizing their spatial coverage.


A global quantification of "normal" sleep schedules using smartphone data.
The influence of the circadian clock on sleep scheduling has been studied extensively in the laboratory; however, the effects of society on sleep remain largely unquantified. We show how a smartphone app that we have developed, ENTRAIN, accurately collects data on sleep habits around the world. Through mathematical modeling and statistics, we find that social pressures weaken and/or conceal biological drives in the evening, leading individuals to delay their bedtime and shorten their sleep. A country's average bedtime, but not average wake time, predicts sleep duration. We further show that mathematical models based on controlled laboratory experiments predict qualitative trends in sunrise, sunset, and light level; however, these effects are attenuated in the real world around bedtime. Additionally, we find that women schedule more sleep than men and that users reporting that they are typically exposed to outdoor light go to sleep earlier and sleep more than those reporting indoor light. Finally, we find that age is the primary determinant of sleep timing, and that age plays an important role in the variability of population-level sleep habits. This work better defines and personalizes "normal" sleep, produces hypotheses for future testing in the laboratory, and suggests important ways to counteract the global sleep crisis.


Frozen-thawed transfer cycles: are they comparable with fresh?
Cryopreservation of zygotes and subsequent thaw and transfer is an established ART treatment. We assessed if success rates frozen-thawed (day 2) zygotes are comparable with the outcome in fresh cycles of treatment. We performed a prospective follow-up and analysis of all frozen (FZT) and fresh cycles of treatment during a 12 months period. One hundred and nineteen patients in the frozen-thawed and 652 in the fresh group had a transfer. The overall thaw-survival rate was 71.7%. Clinical pregnancy rates per thaw and transfer were respectively 15.1% and 21% in the frozen and 29.1% (per transfer) in the fresh group. Implantation rates in fresh and frozen cycles were 16% and 12.3% respectively. The pregnancy loss rate was higher in the FZT group (29% vs. 18.3%). Cryopreservation of good quality zygotes, after fresh transfer offers optimal success rates in subsequent frozen treatment. It also encourages consideration of elective single zygote transfers.


Re-examining the effects of verbal instructional type on early stage motor learning.
The present study investigated the differential effects of analogy and explicit instructions on early stage motor learning and movement in a modified high jump task. Participants were randomly assigned to one of three experimental conditions: analogy, explicit light (reduced informational load), or traditional explicit (large informational load). During the two-day learning phase, participants learned a novel high jump technique based on the 'scissors' style using the instructions for their respective conditions. For the single-day testing phase, participants completed both a retention test and task-relevant pressure test, the latter of which featured a rising high-jump-bar pressure manipulation. Although analogy learners demonstrated slightly more efficient technique and reported fewer technical rules on average, the differences between the conditions were not statistically significant. There were, however, significant differences in joint variability with respect to instructional type, as variability was lowest for the analogy condition during both the learning and testing phases, and as a function of block, as joint variability decreased for all conditions during the learning phase. Findings suggest that reducing the informational volume of explicit instructions may mitigate the deleterious effects on performance previously associated with explicit learning in the literature.


Wildlife habitat connectivity in the changing climate of New York's Hudson Valley.
Maintaining and restoring connectivity are key adaptation strategies for biodiversity conservation under climate change. We present a novel combination of species distribution and connectivity modeling using current and future climate regimes to prioritize connections among populations of 26 rare species in New York's Hudson Valley. We modeled patches for each species for each time period and modeled potential connections among habitat patches by finding the least-cost path for every patch-to-patch connection. Finally, we aggregated these patches and paths to the tax parcel, commonly the primary unit of conservation action. Under future climate regimes, suitable habitat was predicted to contract or appear upslope and farther north. On average, predicted patches were nine times smaller and paths were twice as long under future climate. Parcels within the Hudson Highlands, Shawangunk Ridge, Catskill Mountains, and Harlem Valley had high species overlap, with areas upslope and northward increasing in importance over time. We envision that land managers and conservation planners can use these results to help prioritize parcel-level conservation and management and thus support biodiversity adaptation to climate change.


Inequalities in purchase of mosquito nets and willingness to pay for insecticide-treated nets in Nigeria: challenges for malaria control interventions.
To explore the equity implications of insecticide-treated nets (ITN) distribution programmes that are based on user charges. A questionnaire was used to collect information on previous purchase of untreated nets and hypothetical willingness to pay (WTP) for ITNs from a random sample of householders. A second survey was conducted one month later to collect information on actual purchases of ITNs. An economic status index was used for characterizing inequity. The lower economic status quintiles were less likely to have previously purchased untreated nets and also had a lower hypothetical and actual WTP for ITNs. ITN distribution programmes need to take account of the diversity in WTP for ITNs if they are to ensure equity in access to the nets. This could form part of the overall poverty reduction strategy.


Phase I randomized safety study of twice daily dosing of acidform vaginal gel: candidate antimicrobial contraceptive.
Acidform gel, an acid-buffering product that inactivates spermatozoa, may be an effective topical non-hormonal contraceptive. This study was designed to evaluate the safety of vaginal dosing and effects of Acidform on mucosal immune mediators, antimicrobial properties of genital secretions, and vaginal microbiota. Thirty-six sexually abstinent U.S. women were randomized to apply Acidform or hydroxyethylcellulose (HEC) placebo gel twice daily for 14 consecutive days. Safety was assessed by symptoms and pelvic examination. The impact of gel on mucosal immunity was assessed by quantifying cytokines, chemokines, antimicrobial proteins and antimicrobial activity of genital secretions collected by cervicovaginal lavage (CVL) at screening, 2 hours after gel application, and on days 7, 14 and 21. Vaginal microbiota was characterized at enrollment and day 14 using species-specific quantitative PCR assays. The median vaginal and cervical pH was significantly lower 2 hours after application of Acidform and was associated with an increase in the bactericidal activity of CVL against E. coli. However, 65% of women who received Acidform had at least one local adverse event compared with 11% who received placebo (p = 0.002). While there was no increase in inflammatory cytokines or chemokines, CVL concentrations of lactoferrin and interleukin-1 receptor antagonist (IL-1ra), an anti-inflammatory protein, were significantly lower following Acidform compared to HEC placebo gel application. There were no significant changes in Lactobacillus crispatus or Lactobacillus jensenii in either group but there was a decrease in Gardnerella vaginalis in the Acidform group (p = 0.08). Acidform gel may augment mucosal defense as evidenced by an increase in bactericidal activity of genital secretions against E. coli and a decrease in Gardnerella vaginalis colonization. However, Acidform was associated with more irritation than placebo and lower levels of antimicrobial (lactoferrin) and anti-inflammatory (IL-1ra) proteins. These findings indicate the need for additional safety studies of this candidate non-hormonal contraceptive. ClinicalTrials.gov NCT00850837.


Association of phosphorylated epidermal growth factor receptor with survival in patients with locally advanced non-small cell lung cancer treated with radiotherapy.
The epidermal growth factor receptor (EGFR) and its downstream effector kinases are thought to have important roles in lung cancer cell proliferation and response to treatment. In a prospective cohort of patients with locally advanced non-small cell lung cancer (NSCLC) undergoing high-dose radiotherapy with or without chemotherapy, we examined by immunohistochemistry (IHC) the tumor levels of EGFR and phosphorylated/ activated-EGFR (P-EGFR), P-Erk, P-Akt, P-Stat3, and the cell cycle marker Ki67. We examined the relationships among marker expression, at the plasma membrane (M), cytoplasm (C) and nucleus, as well as the radiologic tumor response, and overall survival (OS). Fifty patients were recruited in this study. Their median survival was 18.3 months. No correlation was detected between total EGFR and P-EGFR levels in any subcellular compartment. M P-EGFR correlated positively with C P-Akt (p = 0.01). C P-Erk correlated negatively with radiologic response (p = 0.022), and on univariate regression analysis, this approached significance (p = 0.059). M and C P-EGFR correlated negatively with OS (p = 0.020), and on univariate analysis higher M P-EGFR predicted for poor OS (p = 0.001). Patients with high M P-EGFR levels had median survival of 7.8 months versus 17.7 months for patients with low M P-EGFR. Our results support a role of activated M P-EGFR, (but not total EGFR), as a predictor of OS in locally advanced-NSCLC. It is suggested that detailed evaluation of the subcellular distribution and activation state of EGFR and its downstream effectors is required to unravel the predictive value of these markers in NSCLC.


Factors influencing experience in crowds - The organiser perspective.
Crowds are a commonplace encounter but the experience for participants can be highly variable. Crowds are complex sociotechnical phenomenon, affected by many interacting factors. Little is known, however, about how those responsible for organising crowd situations approach their responsibilities. This study conducted semi-structured interviews (n = 41) with organisers responsible for different aspects of the design, planning, management and operations of events and other crowd situations. The objective was to understand organisers' priorities, along with the consideration given to the experience of crowd participants. The interviews revealed that organisers generally prioritised finance, security and health and safety aspects, whilst giving limited explicit attention to other important factors that affect participant experience. Organisers tended to approach their planning and decisions on the basis of their own experience and judgement, without accessing training or reference to guidance. It is suggested that the non-use of guidance is in part due to problems with the guidance currently available, both its content and its form. The organisers of infrequent or small-scale events have the greatest knowledge and experience gap. It is concluded that in order to achieve a consistent, high quality experience for crowd participants, there needs to be improved understanding among organisers of the complexity of crowds and the multiple factors influencing participant experience. Guidance and tools need to be usable and tailored to organisers' requirements. Organisers of infrequent or small-scale events are especially in need of support.


Blood pressure parameters and carotid intraplaque hemorrhage as measured by magnetic resonance imaging: The Rotterdam Study.
Intraplaque hemorrhage (IPH) is a characteristic of the vulnerable atherosclerotic plaque that has been associated with ischemic stroke. Not much is known about determinants of IPH. We studied whether blood pressure parameters are associated with presence of IPH. Within the framework of a prospective population-based cohort study, The Rotterdam Study, the carotid arteries of 1006 healthy participants ≥45 years and with intima-media thickening (≥2.5 mm) on ultrasound were imaged with a 1.5-T magnetic resonance imaging scanner. IPH was defined as a hyperintense signal on a 3D-T1w-GRE magnetic resonance sequence. Generalized estimation equation analysis, adjusted for age, sex, carotid wall thickness, and cardiovascular risk factors, was used to assess the association between blood pressure parameters and IPH. Magnetic resonance imaging of the carotid arteries revealed presence of IPH in 444 of 1860 plaques (24%). Systolic blood pressure and pulse pressure (PP) were significantly associated with IPH after adjustment for age and sex. In multivariate analysis, PP yielded the strongest association, with an odds ratio per SD increase in PP of 1.22 (95% CI, 1.07-1.40). The odds ratio per SD for systolic blood pressure was 1.13 (0.99-1.28). Only PP remained significant after additional adjustment for other blood pressure components. The combination of smoking and isolated systolic hypertension was associated with 2.5 times increased risk of IPH (1.2-5.2). In conclusion, PP was the strongest determinant of IPH independent of cardiovascular risk factors and other blood pressure components. The association between pulsatile flow and IPH may provide novel insights in the development of the vulnerable plaque.


Bee Venom Phospholipase A2, a Good "Chauffeur" for Delivering Tumor Antigen to the MHC I and MHC II Peptide-Loading Compartments of the Dendritic Cells: The Case of NY-ESO-1.
Bee venom phospholipase A2 (bvPLA2) is a small, 15kDa enzyme which hydrolyses many phospholipids through interfacial binding. The mutated bvPLA2H34Q (bvPLA2m), in which histidine-34 is replaced by glutamine, is not catalytically active. This protein has been shown to be a suitable membrane anchor and has been suggested as a suitable tumor-antigen vector for the development of novel dendritic cell-based vaccines. To confirm this feature, in this study the fusion protein PNY, composed of NY-ESO-1(NY(s)) fused to the C-terminus of bvPLA2m, was engineered. bvPLA2m enhanced the binding of NY(s) to the membrane of human monocyte-derived dendritic cells (DCs) and, once taken up by the cells, the antigen fused to the vector was directed to both MHC I and MHC II peptide-loading compartments. bvPLA2m was shown to increase the cross-presentation of the NY(s)-derived, restricted HLA-A*02 peptide, NY-ESO-1157-165(NY157-165), at the T1 cell surface. DCs loaded with the fusion protein induced cross-priming of NY(s)-specific CD8 + T-cells with greater efficiency than DCs loaded with NY(s). Sixty-five percent of these NY(s)-specific CD8+ T-cell lines could also be activated with the DCs pulsed with the peptide, NY157-165. Of these CD8+ T-cell lines, two were able to recognize the human melanoma cell line, SK-MEL-37, in a context of HLA-A*02. Only a small number of bvPLA2m CD8+ T-cell lines were induced, indicating the low immunogenicity of the protein. It was concluded that bvPLA2m can be used as a membrane-binding vector to promote MHC class II peptide presentation and MHC class I peptide cross-presentation. Such a system can, therefore, be tested for the preparation of cell-based vaccines.


Anatomy, histochemistry, and immunohistochemistry of the olfactory subsystems in mice.
The four regions of the murine nasal cavity featuring olfactory neurons were studied anatomically and by labeling with lectins and relevant antibodies with a view to establishing criteria for the identification of olfactory subsystems that are readily applicable to other mammals. In the main olfactory epithelium and the septal organ the olfactory sensory neurons (OSNs) are embedded in quasi-stratified columnar epithelium; vomeronasal OSNs are embedded in epithelium lining the medial interior wall of the vomeronasal duct and do not make contact with the mucosa of the main nasal cavity; and in Grüneberg's ganglion a small isolated population of OSNs lies adjacent to, but not within, the epithelium. With the exception of Grüneberg's ganglion, all the tissues expressing olfactory marker protein (OMP) (the above four nasal territories, the vomeronasal and main olfactory nerves, and the main and accessory olfactory bulbs) are also labeled by Lycopersicum esculentum agglutinin, while Ulex europaeus agglutinin I labels all and only tissues expressing Gαi2 (the apical sensory neurons of the vomeronasal organ, their axons, and their glomerular destinations in the anterior accessory olfactory bulb). These staining patterns of UEA-I and LEA may facilitate the characterization of olfactory anatomy in other species. A 710-section atlas of the anatomy of the murine nasal cavity has been made available on line.


Food sources of fat may clarify the inconsistent role of dietary fat intake for incidence of type 2 diabetes.
Dietary fats could affect glucose metabolism and obesity development and, thereby, may have a crucial role in the cause of type 2 diabetes (T2D). Studies indicated that replacing saturated with unsaturated fats might be favorable, and plant foods might be a better choice than animal foods. Nevertheless, epidemiologic studies suggested that dairy foods are protective. We hypothesized that, by examining dietary fat and its food sources classified according to fat type and fat content, some clarification regarding the role of dietary fat in T2D incidence could be provided. A total of 26,930 individuals (61% women), aged 45-74 y, from the Malmö Diet and Cancer cohort were included in the study. Dietary data were collected by using a modified diet-history method. During 14 y of follow-up, 2860 incident T2D cases were identified. Total intake of high-fat dairy products (regular-fat alternatives) was inversely associated with incident T2D (HR for highest compared with lowest quintiles: 0.77; 95% CI: 0.68, 0.87; P-trend < 0.001). Most robust inverse associations were seen for intakes of cream and high-fat fermented milk (P-trend < 0.01) and for cheese in women (P-trend = 0.02). High intake of low-fat dairy products was associated with increased risk, but this association disappeared when low- and high-fat dairy were mutually adjusted (P-trend = 0.18). Intakes of both high-fat meat (P-trend = 0.04) and low-fat meat (P-trend < 0.001) were associated with increased risk. Finally, we did not observe significant association between total dietary fat content and T2D (P-trend = 0.24), but intakes of saturated fatty acids with 4-10 carbons, lauric acid (12:0), and myristic acid (14:0) were associated with decreased risk (P-trend < 0.01). Decreased T2D risk at high intake of high- but not of low-fat dairy products suggests that dairy fat partly could have contributed to previously observed protective associations between dairy intake and T2D. Meat intake was associated with increased risk independently of the fat content.


Imaging density disturbances in water with a 41.3-attosecond time resolution.
We show that the momentum flexibility of inelastic x-ray scattering may be exploited to invert its loss function, allowing real time imaging of density disturbances in a medium. We show the disturbance arising from a point source in liquid water, with a resolution of 41.3 attoseconds (4.13 x 10(-17) s) and 1.27 A (1.27 x 10(-8) cm). This result is used to determine the structure of the electron cloud around a photoexcited chromophore in solution, as well as the wake generated in water by a 9 MeV gold ion. We draw an analogy with pump-probe techniques and suggest that energy-loss scattering may be applied more generally to the study of attosecond phenomena.


Oncofertility: Knowledge, Attitudes, and Barriers Among Indian Oncologists and Gynecologists.
Purpose: Recommendations from the American Society of Clinical Oncology (ASCO) emphasize the critical need to understand current trends in fertility preservation (FP) among the two sets of primary health care providers involved in oncofertility: the oncologists and the gynecologists. This study is aimed at understanding the health care providers' knowledge, attitudes, and barriers in oncofertility across India. Methods: An 18-item oncofertility survey was designed and directed to 77 oncologists and 214 gynecologists across India. The responses were analyzed by using descriptive statistical methods, and the oncofertility trends between the two groups were studied. Results: The total response rate was 34%, with 49 of 214 oncologists (23%) and 49 of 77 gynecologists (64%) participating in the survey. The awareness of ASCO FP guidelines among oncologists and gynecologists was 53% and 59.5%, respectively. About 48% of oncologists felt knowledgeable about sperm banking, whereas 52% knew about oocyte freezing but not about other options. On the other hand, among gynecologists, 38% reported inadequate knowledge of testicular or ovarian tissue cryopreservation. About 85% of oncologists reported routine referral of cancer diagnosed patients for FP, whereas 75% of gynecologists reported routine FP discussion with patients. Health care providers from both groups perceived the major barriers in oncofertility to be, "financial burden on the patient" (73%-86%) and, "lack of patient awareness" (71%-79.5%). Conclusion: Effective collaboration between oncologists and gynecologists is essential to establish a successful FP program. Economic burden on the patient and lack of patient and physician awareness are limiting factors that need to be overcome.


Systemic and cell type-specific gene expression patterns in scleroderma skin.
We used DNA microarrays representing >12,000 human genes to characterize gene expression patterns in skin biopsies from individuals with a diagnosis of systemic sclerosis with diffuse scleroderma. We found consistent differences in the patterns of gene expression between skin biopsies from individuals with scleroderma and those from normal, unaffected individuals. The biopsies from affected individuals showed nearly indistinguishable patterns of gene expression in clinically affected and clinically unaffected tissue, even though these were clearly distinguishable from the patterns found in similar tissue from unaffected individuals. Genes characteristically expressed in endothelial cells, B lymphocytes, and fibroblasts showed differential expression between scleroderma and normal biopsies. Analysis of lymphocyte populations in scleroderma skin biopsies by immunohistochemistry suggest the B lymphocyte signature observed on our arrays is from CD20+ B cells. These results provide evidence that scleroderma has systemic manifestations that affect multiple cell types and suggests genes that could be used as potential markers for the disease.


[Clinicalpathological characteristics of Lynch syndrome related epithelial ovarian cancer].
To explore the clinical pathological characteristics of Lynch syndrome associated ovarian cancer. Totally 260 cases ovarian cancer patients were admitted to Tianjin Medical University General Hospital during Jan. 2004 and Jan. 2011, among which 10 patients (LS group) belonged to Lynch syndrome associated ovarian cancer according to Amsterdam II criteria. One hundred ovarian cancer patients without any family cancer history were enrolled randomizely as control group (sporadic group). Lynch syndrome associated ovarian cancer accounted for 3.8% (10/260), the incidence rate of ovarian cancer for female family members of Lynch syndrome was 8.7% (10/115). Mean age at time of diagnosis in LS group was (46 ± 7) years, significantly earlier than that in sporadic group [(56 ± 11) years, P < 0.05]. There was no statistical difference between two groups in histological type or International Federation of Gynecology and Obstetrics (FIGO) stage (P > 0.05). Most of the tissue differentiation in LS group were well or moderate differentiated, there was statistical difference between the two groups (9/10 vs. 55%, P < 0.05). The 3-year and 5-year survival rate in LS group were 87.5% and 52.5% respectively, compared with 55.4%and 22.7% in sporadic group (all P < 0.05). Compared with sporadic ovarian cancer, Lynch syndrome associated ovarian cancer is more likely present as the clinical pathological characteristics of early age of onset, serous adenocarcinoma, lower grade and better prognosis.


Dynamic 3-dimensional stress cardiac magnetic resonance perfusion imaging: detection of coronary artery disease and volumetry of myocardial hypoenhancement before and after coronary stenting.
The aim of this study was to establish a new, dynamic 3-dimensional cardiac magnetic resonance (3D-CMR) perfusion scan technique exploiting data correlation in k-space and time with sensitivity-encoding and to determine its value for the detection of coronary artery disease (CAD) and volumetry of myocardial hypoenhancement (VOLUME(hypo)) before and after percutaneous coronary stenting. Dynamic 3D-CMR perfusion imaging might improve detection of myocardial perfusion deficits and could facilitate direct volumetry of myocardial hypoenhancement. In 146 patients with known or suspected CAD, a 3.0-T CMR examination was performed including cine imaging, 3D-CMR perfusion under adenosine stress and at rest followed by delayed enhancement imaging. Quantitative invasive coronary angiography defined significant CAD (≥ 50% luminal narrowing). Forty-eight patients underwent an identical repeat CMR examination after percutaneous stenting of at least 1 coronary lesion. The 3D-CMR perfusion scans were visually classified as pathologic if ≥ 1 segment showed an inducible perfusion deficit in the absence of delayed enhancement. The VOLUME(hypo) was measured by segmentation of the area of inducible hypoenhancement and normalized to left-ventricular myocardial volume (%VOLUME(hypo)). The 3D-CMR perfusion resulted in a sensitivity, specificity, and diagnostic accuracy of 91.7%, 74.3%, and 82.9%, respectively. Before and after coronary stenting, %VOLUME(hypo) averaged to 14.2 ± 9.5% and 3.2 ± 5.2%, respectively, with a relative VOLUME(hypo) reduction of 79.4 ± 25.4%. Intrareader and inter-reader reproducibility of VOLUME(hypo) measurements was high (Lin's concordance correlation coefficient, 0.96 and 0.96, respectively). The 3D-CMR stress perfusion provided high image quality and high diagnostic accuracy for the detection of significant CAD. The VOLUME(hypo) measurements were highly reproducible and allowed for the assessment of the treatment effect achievable by percutaneous coronary stenting.


The effects of road traffic and aircraft noise exposure on children's episodic memory: the RANCH project.
Previous studies have found that chronic exposure to aircraft noise has a negative effect on children's performance on tests of episodic memory. The present study extended the design of earlier studies in three ways: firstly, by examining the effects of two noise sources, aircraft and road traffic, secondly, by examining exposure-effect relationships, and thirdly, by carrying out parallel field studies in three European countries, allowing cross-country comparisons to be made. A total of 2844 children aged between 8 years 10 months and 12 years 10 months (mean age 10 years 6 months) completed classroom-based tests of cued recall, recognition memory and prospective memory. Questionnaires were also completed by the children and their parents in order to provide information about socioeconomic context. Multilevel modeling analysis revealed aircraft noise to be associated with an impairment of recognition memory in a linear exposure-effect relationship. The analysis also found road traffic noise to be associated with improved performance on cued recall in a linear exposure-effect relationship. No significant association was found between exposure to aircraft noise and cued recall or prospective memory. Likewise, no significant association was found between road traffic noise and recognition or prospective memory. Taken together, these findings indicate that exposure to aircraft noise and road traffic noise can impact on certain aspects of children's episodic memory.


Gregarine Oocyst Production is a Function of Gametocyst Size.
Gregarine transmission depends upon the environmental encounter rate between viable infective oocysts and suitable hosts. Many factors determine the abundance and distribution of gregarine oocysts in the environment, but the primary factors are oocyst distribution, environmental persistence, and production rate. Prior studies have demonstrated factors affecting oocyst distribution and environmental persistence, but oocyst production rate is poorly understood. This study addresses the effects of gametocyst size on oocyst production. For each of 3 gregarine species, gametocyst size was determined, and the subsequent oocyst production of each gametocyst was quantified. Gregarine species with larger gametocysts produced more oocysts per gametocyst than species with smaller gametocysts. Likewise, within species, larger gametocysts produced more oocysts. The effect was stronger in larger gregarine species, probably as a reflection of the lower overall range of gametocyst size in the smaller gregarine species.


A randomized controlled trial to compare pregabalin with gabapentin for postoperative pain in abdominal hysterectomy.
Pregabalin is a potent ligand for alpha-2-delta subunit of voltage-gated calcium channels in the central nervous system, which exhibits potent anticonvulsant, analgesic and anxiolytic activity. The pharmacological activity of pregabalin is similar to that of gabapentin and shows possible advantages. Although it shows analgesic efficacy against neuropathic pain, very limited evidence supports its postoperative analgesic efficacy. We investigated its analgesic efficacy in patients experiencing acute pain after abdominal hysterectomy and compared it with gabapentin and placebo. A randomized, double-blind, placebo-controlled study was conducted in 90 women undergoing abdominal hysterectomy who were anaesthetized in a standardized fashion. Patients received 300 mg pregabalin, 900 mg gabapentin or placebo, 1-2 hours prior to surgery. Postoperative analgesia was administered at visual analogue scale (VAS) ≥3. The primary outcome was analgesic consumption over 24 hours and patients were followed for pain scores, time to rescue analgesia and side effects as secondary outcomes. The diclofenac consumption was statistically significant between pregabalin and control groups, and gabapentin and control groups; however, pregabalin and gabapentin groups were comparable. Moreover, the consumption of tramadol was statistically significant among all the groups. Patients in pregabalin and gabapentin groups had lower pain scores in the initial hour of recovery. However, pain scores were subsequently similar in all the groups. Time to first request for analgesia was longer in pregabalin group followed by gabapentin and control groups. A single dose of 300 mg pregabalin given 1-2 hours prior to surgery is superior to 900 mg gabapentin and placebo after abdominal hysterectomy. Both the drugs are better than placebo.


Natural epitope variants of the hepatitis C virus impair cytotoxic T lymphocyte activity.
To understand how interactions between hepatitis C virus (HCV) and the host's immune system might lead to viral persistence or effective elimination of HCV. Nucleotides 3519-3935 of the non-structural 3 (NS3) region were amplified by using reverse transcription polymerase chain reaction (PCR). PCR products of the HCV NS3 regions were integrated into a PCR((R)) T7TOPO((R)) TA vector and then sequenced in both directions using an automated DNA sequencer. Relative major histocompatibility complex binding levels of wild-type and variant peptides were performed by fluorescence polarization-based peptide competition assays. Peptides with wild type and variant sequences of NS3 were synthesized locally using F-moc chemistry and purified by high-performance liquid chromatography. Specific cytotoxic T lymphocytes (CTLs) clones toward HCV NS3 wild-type peptides were generated through limiting dilution cloning. The CTL clones specifically recognizing HCV NS3 wild-type peptides were tested by tetramer staining and flow cytometry. Cytolytic activity of CTL clones was measured using target cells labeled with the fluorescence enhancing ligand, DELFIA EuTDA. The pattern of natural variants within three human leukocyte antigen (HLA)-A2-restricted NS3 epitopes has been examined in one patient with chronic HCV infection at 12, 28 and 63 mo post-infection. Results obtained may provide convincing evidence of immune selection pressure for all epitopes investigated. Statistical analysis of the extensive sequence variation found within these NS3 epitopes favors a Darwinian selection model of variant viruses. Mutations within the epitopes coincided with the decline of CTL responses, and peptide-binding studies suggested a significant impact of the mutation on T cell recognition rather than peptide presentation by HLA molecules. While most variants were either not recognized or elicited low responses, such could antagonize CTL responses to target cells pulsed with wild-type peptides. Cross-recognition of CTL epitopes from wild-type and naturally-occurring HCV variants may lead to impaired immune responses and ultimately contribute to viral persistence.


Study protocol for the SMART2D adaptive implementation trial: a cluster randomised trial comparing facility-only care with integrated facility and community care to improve type 2 diabetes outcomes in Uganda, South Africa and Sweden.
Type 2 diabetes (T2D) is increasingly contributing to the global burden of disease. Health systems in most parts of the world are struggling to diagnose and manage T2D, especially in low-income and middle-income countries, and among disadvantaged populations in high-income countries. The aim of this study is to determine the added benefit of community interventions onto health facility interventions, towards glycaemic control among persons with diabetes, and towards reduction in plasma glucose among persons with prediabetes. An adaptive implementation cluster randomised trial is being implemented in two rural districts in Uganda with three clusters per study arm, in an urban township in South Africa with one cluster per study arm, and in socially disadvantaged suburbs in Stockholm, Sweden with one cluster per study arm. Clusters are communities within the catchment areas of participating primary healthcare facilities. There are two study arms comprising a facility plus community interventions arm and a facility-only interventions arm. Uganda has a third arm comprising usual care. Intervention strategies focus on organisation of care, linkage between health facility and the community, and strengthening patient role in self-management, community mobilisation and a supportive environment. Among T2D participants, the primary outcome is controlled plasma glucose; whereas among prediabetes participants the primary outcome is reduction in plasma glucose. The study has received approval in Uganda from the Higher Degrees, Research and Ethics Committee of Makerere University School of Public Health and from the Uganda National Council for Science and Technology; in South Africa from the Biomedical Science Research Ethics Committee of the University of the Western Cape; and in Sweden from the Regional Ethical Board in Stockholm. Findings will be disseminated through peer-reviewed publications and scientific meetings. ISRCTN11913581; Pre-results.


From carotenoid intake to carotenoid blood and tissue concentrations - implications for dietary intake recommendations.
There is uncertainty regarding carotenoid intake recommendations, because positive and negative health effects have been found or are correlated with carotenoid intake and tissue levels (including blood, adipose tissue, and the macula), depending on the type of study (epidemiological vs intervention), the dose (physiological vs supraphysiological) and the matrix (foods vs supplements, isolated or used in combination). All these factors, combined with interindividual response variations (eg, depending on age, sex, disease state, genetic makeup), make the relationship between carotenoid intake and their blood/tissue concentrations often unclear and highly variable. Although blood total carotenoid concentrations <1000 nmol/L have been related to increased chronic disease risk, no dietary reference intakes (DRIs) exist. Although high total plasma/serum carotenoid concentrations of up to 7500 nmol/L are achievable after supplementation, a plateauing effect for higher doses and prolonged intake is apparent. In this review and position paper, the current knowledge on carotenoids in serum/plasma and tissues and their relationship to dietary intake and health status is summarized with the aim of proposing suggestions for a "normal," safe, and desirable range of concentrations that presumably are beneficial for health. Existing recommendations are likewise evaluated and practical dietary suggestions are included.


Modulation of epidermal differentiation by epiligrin and integrin alpha 3 beta 1.
We previously reported that integrin alpha 3 beta 1 mediates epidermal intercellular adhesion as well as cell-substrate adhesion. P1B5, an anti-alpha 3 beta 1 specific monoclonal antibody, is a potent in vitro trigger of epidermal cell-cell adhesion and an inhibitor of cell-substrate adhesion. We now show that P1B5 specifically induces the intercellular localization of integrins alpha 2 beta 1 and alpha 3 beta 1, consistent with its role in inducing intercellular adhesion via these two integrins. P1F2, another anti-alpha 3 beta 1 antibody, does not induce either intercellular adhesion or intercellular accumulation of alpha 3 beta 1 and alpha 2 beta 1. Growth of epidermal cells in high calcium, known to induce epidermal differentiation, also induces intercellular accumulation of alpha 3 beta 1 and alpha 2 beta 1 and increased cell-cell adhesion. We therefore asked whether P1B5 treatment induces epidermal differentiation. P1B5 treatment induces changes consistent with epidermal differentiation, including increased involucrin expression, stratification, and production of squames. P1F2 treatment has none of these effects. In vivo, epidermal basal cells are in close contact with the epithelial basement membrane component epiligrin. Growth of keratinocytes on purified epiligrin but not other matrix components specifically reduces involucrin expression by P1B5-treated keratinocytes. These results suggest that integrin alpha 3 beta 1 has a unique role in epidermal differentiation, that the epitope recognized by P1B5 is involved in triggering this differentiation, and that keratinocyte adhesion to epiligrin inhibits alpha 3 beta 1-mediated differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)


Large objects reconstructed from computer-generated holograms.
In digital holography the first step is the determination of the complex amplitude in the hologram. Large objects can be synthesized from tilted planar segments. In the calculation step, configurations of inclined object and inclined hologram plane are considered. The Fresnel transformation is generalized, and its implementation is discussed. Optical reconstructions show the feasibility.


Two new constraints for the cumulant matrix.
We suggest new strict constraints that the two-particle cumulant matrix should fulfill. The constraints are obtained from the decomposition of ⟨Ŝ(2)⟩, previously developed in our laboratory, and the vanishing number of electrons shared by two non-interacting fragments. The conditions impose stringent constraints into the cumulant structure without any need to perform an orbital optimization procedure thus carrying very small or no computational effort. These constraints are tested on the series of Piris natural orbital functionals (PNOF), which are among the most accurate ones available in the literature. Interestingly, even though all PNOF cumulants ensure correct overall ⟨Ŝ(2)⟩ values, none of them is consistent with the local spin structure of systems that dissociate more than one pair of electrons. A careful analysis of the local spin components reveals the most important missing contributions in the cumulant expression thus suggesting a means to improve PNOF5. The constraints provide an inexpensive tool for the construction and testing of cumulant structures that complement previously known conditions such as the N-representability or the square of the total spin angular momentum, ⟨Ŝ(2)⟩.


PRMT4 blocks myeloid differentiation by assembling a methyl-RUNX1-dependent repressor complex.
Defining the role of epigenetic regulators in hematopoiesis has become critically important, because recurrent mutations or aberrant expression of these genes has been identified in both myeloid and lymphoid hematological malignancies. We found that PRMT4, a type I arginine methyltransferase whose function in normal and malignant hematopoiesis is unknown, is overexpressed in acute myelogenous leukemia patient samples. Overexpression of PRMT4 blocks the myeloid differentiation of human stem/progenitor cells (HSPCs), whereas its knockdown is sufficient to induce myeloid differentiation of HSPCs. We demonstrated that PRMT4 represses the expression of miR-223 in HSPCs via the methylation of RUNX1, which triggers the assembly of a multiprotein repressor complex that includes DPF2. As part of the feedback loop, PRMT4 expression is repressed posttranscriptionally by miR-223. Depletion of PRMT4 results in differentiation of myeloid leukemia cells in vitro and their decreased proliferation in vivo. Thus, targeting PRMT4 holds potential as a novel therapy for acute myelogenous leukemia.


Facile fabrication of highly controllable gating systems based on the combination of inverse opal structure and dynamic covalent chemistry.
A three-dimensional (3D) inverse opal with periodic and porous structures has shown great potential for applications not only in optics and optoelectronics, but also in functional membranes. In this work, the benzaldehyde group was initially introduced into a 3D nanoporous inverse opal, serving as a platform for fabricating functional membranes. By employing the dynamic covalent approach, a highly controllable gating system was facilely fabricated to achieve modulable and reversible transport features. It was found that the physical/chemical properties and pore size of the gating system could easily be regulated through post-modification with amines. As a demonstration, the gated nanopores were modified with three kinds of amines to control the wettability, surface charge and nanopore size which in turn was exploited to achieve selective mass transport, including hydrophobic molecules, cations and anions, and the transport with respect to the physical steric hindrance. In particular, the gating system showed extraordinary reversibility and could recover to its pristine state by simply changing pH values. Due to the unlimited variety provided by the Schiff base reaction, the inverse opal described here exhibits a significant extendibility and could be easily post-modified with stimuli-responsive molecules for special purposes. Furthermore, this work can be extended to employ other dynamic covalent routes, for example Diels-Alder, ester exchange and disulfide exchange-based routes.


Amelioration of behavioral aberrations and oxidative markers by green tea extract in valproate induced autism in animals.
Exposure to toxicants prenatally and postnatally could have deleterious consequences on the offspring. Postnatal exposure to valproate in mice pups is capable of inducing experimental autism resulting in neurobehavioral aberrations. Consumption of green tea has been associated with neuronal protection against the impact of toxicants. We investigated the role of green tea extract in reversing cardinal behavioral changes and aberrations in oxidative stress induced by valproate exposure. Young mice of both genders received a single dose of valproate (400mg/kg subcutaneously) on postnatal day 14 followed by a daily dose of green tea extract (75 and 300mg/kg) orally up to postnatal day 40. Mice pups were subjected to behavioral testing to assess motor co-ordination, nociceptive response, locomotion, anxiety, exploratory activity and cognition on various postnatal days up to postnatal day 40. At the end of behavioral testing, blood was withdrawn from the retro orbital plexus for the estimation of lipid peroxides. Animals were sacrificed on postnatal day 41 and whole brain was subjected to histopathological examination. Our studies revealed a significant improvement in behavioral assessments particularly with 300mg/kg of green tea extract. Formation of markers of oxidative stress was reduced at both dose levels. Histological findings confirm the neuroprotective effect of green tea at a dose of 300mg/kg. In conclusion it can be stated that green tea exerts neuronal cytoprotective action possibly due to anti-oxidant action and could be efficacious in the management of autism.


Frequent detection of Streptococcus tigurinus in the human oral microbial flora by a specific 16S rRNA gene real-time TaqMan PCR.
Many bacteria causing systemic invasive infections originate from the oral cavity by entering the bloodstream. Recently, a novel pathogenic bacterium, Streptococcus tigurinus, was identified as causative agent of infective endocarditis, spondylodiscitis and meningitis. In this study, we sought to determine the prevalence of S. tigurinus in the human oral microbial flora and analyzed its association with periodontal disease or health. We developed a diagnostic highly sensitive and specific real-time TaqMan PCR assay for detection of S. tigurinus in clinical samples, based on the 16S rRNA gene. We analyzed saliva samples and subgingival plaque samples of a periodontally healthy control group (n = 26) and a periodontitis group (n = 25). Overall, S. tigurinus was detected in 27 (53%) out of 51 patients. There is no significant difference of the frequency of S. tigurinus detection by RT-PCR in the saliva and dental plaque samples in the two groups: in the control group, 14 (54%) out of 26 individuals had S. tigurinus either in the saliva samples and/or in the plaque samples; and in the periodontitis group, 13 (52%) out of 25 patients had S. tigurinus in the mouth samples, respectively (P = 0.895). The consumption of nicotine was no determining factor. Although S. tigurinus was a frequently detected species of the human oral microbial flora, it was not associated with periodontal disease. Further investigations are required to determine whether S. tigurinus is a commensal or an opportunistic oral pathogen with a potential for development of invasive infections.


Quasi-periodic transformations of nonlocal spatial solitons.
We study quasi-periodic transformations between nonlocal spatial solitons of different symmetries triggered by modulational instability and resembling a self-induced mode converter. Transformation dynamics of solitons with zero angular momentum, e.g. the quadrupole-type soliton, reveal the equidistant spectrum of spatial field oscillations typical for the breather-type solutions. In contrast, the transformations of nonlocal solitons carrying orbital angular momentum, such as 2x3 soliton matrix, are accompanied by their spiralling and corresponding spectra of field oscillations show mixing of three characteristic spatial frequencies.


Expression and regulation of BCL-2 family genes in human adrenocortical adenomas in comparison with adrenal hyperplasia of Cushing's disease.
The significance of increases in the expression of apoptosis-suppressing genes such as bcl-2 and mcl-1/EAT in human adrenal tumors has not yet been fully elucidated. Furthermore the roles of these genes in cell proliferation may involve interaction with steroidogenesis in the tumors via intracellular second messengers. Cyclic AMP (cAMP) caused human adrenocortical H295R cells to overexpress hCYP17 resulting in hypersecretion of cortisol. At the same time, however, expression of bcl-2, which has a cAMP response element (CRE), was not affected. Furthermore, in vivo Bcl-2 protein analysis showed its down-regulation in adrenal hyperplasia of Cushing's disease despite ACTH stimulation. Exogenous addition of glucocorticoid did not affect the expression of bcl-2 family genes. Expressions of Mcl-1/EAT and Bax did not differ markedly among human adrenal glands affected by various pathologies. In conclusion the down-regulation of Bcl-2 in Cushing's disease did not agree with no induction of this gene by cAMP in H295R cells, suggesting that expression of Bcl-2 protein was not regulated mainly by cAMP-protein kinase (PKA) pathways in human adrenal hyperplasia.


Free-tissue transfer for limb salvage in Purpura fulminans.
A series of 13 patients is described to demonstrate the experience of the authors with free-tissue transfer for limb salvage in patients with purpura fulminans. A total of seven free-flap procedures were performed, with a loss of flap in one patient. The flaps were used for lower-extremity salvage in six patients and for upper-extremity salvage in one. Purpura fulminans is a devastating illness caused by endotoxin-producing bacteria such as meningococcus and pneumococcus. Clotting derangements and systemic vasculitis often lead to widespread tissue necrosis in the extremities. Local tissue is usually not available to cover vital structures in these complex wounds. In these situations, free-tissue transfer is necessary to achieve limb salvage. Microsurgical reconstruction in patients with purpura fulminans is a formidable challenge. Because of high platelet counts and systemic vasculitis, successful microvascular anastomosis is difficult. Abnormally high platelet counts persist well into the subacute and chronic phases of the illness. Pretreatment with antiplatelet agents before microvascular surgery may be beneficial. The systemic nature of the vascular injury does not permit microvascular anastomosis to be performed outside the "zone of injury." Extensive vascular exposure, even at a great distance from the wound, does not reveal a disease-free vessel. The friable intima is difficult to manage with a standard end-to-side anastomosis, but conversion to end-to-end anastomosis may salvage free-tissue transfers in cases in which intimal damage is too severe to sustain a patent anastomosis. Patients often have peripheral neuropathies caused by the underlying disease; however, this resolves with time and is not a contraindication to limb salvage.


Multi-component assessment of worker exposures in a copper refinery. Part 1. Environmental monitoring.
The exposure characterisation described in this paper for 135 copper refinery workers (45 females, 90 males) focuses on the concentrations of copper, nickel and other trace elements in the inhalable aerosol fractions, as well as in the water-soluble and water-insoluble subfractions. Some information is also provided on the thoracic and respirable aerosol fractions. Further, results are presented for volatile hydrides of arsenic and selenium released in the copper purification steps of the electrorefining process. For the pyrometallurgical operations, a comparison of the geometric means for the inhalable aerosol fraction indicated that water-soluble copper levels were on average 19-fold higher compared to nickel (p < 0.001) and a significant association was evident between them (r = 0.87, p < 0.001); for the insoluble subfraction, the copper : nickel ratio was 12.5 (p < 0.001) and the inter-element correlation had r = 0.98 and p < 0.001. Although for the electrorefinery workers the relative inhalable concentrations of copper and nickel were not significantly different (p > 0.05), the corresponding inter-element associations were: slope of 7.7, r= 0.54, p < or =0.001 for the water-soluble subfraction and slope of 1.3, r = 0.71 and p < or =0.001 for the water-insoluble subfraction. On average, a good proportion of the inhalable copper and nickel were found in the thoracic (40%) and respirable (20%) aerosol fractions. Cobalt air concentrations were generally low with geometric means and 95% confidence intervals of 3.1 (2.4-4.2)microg m(-3) (pyrometallurgical workers) and 0.3 (0.4-0.5) microg m(-3)(electrorefinery workers). Similarly, the maximum concentrations of cadmium and lead were low, respectively 4 and 25 microg m(-3). Of the hydrides, tellurium and antimony could not be detected, but for the arsenic (arsine) and selenium hydrides measurable exposure occurred for almost all electrorefinery workers, although the levels were generally low at 0.2 microg m(-3).


Independent prognostic variables in acute myeloid leukaemia.
Acute myeloid leukaemia (AML) is one of the most common haematological malignancies and is increasing in frequency due to an ageing population. Whilst remission will be achieved in up to 80% of those receiving intensive chemotherapy, the main variables precluding cure are the treatment-related mortality and relapse rates. Decisions on intensification, de-escalation and allografting rely on the ability to divide an apparently homogeneous group according to risk. A wide range of clinical, cytogenetic and molecular variables may be used to inform this task. Cytogenetic and molecular characterisation has already identified subgroups, such as acute promyelocytic leukaemia (APL) with t(15;17)/PML-RARA and AML with FLT3 mutation for which targeted therapies are available, and further molecularly defined groups who may be potential candidates for this approach are likely to be identified in the future. This review examines the range of established clinical and diagnostic parameters that should be used in assessing prognosis for a patient with AML and looks ahead to an expanding repertoire of potential variables that are currently under evaluation.


Impact on seizure control of surgical resection or radiosurgery for cerebral arteriovenous malformations.
Seizures are a common presenting symptom of arteriovenous malformations (AVMs). However, the impact of treatment modality on seizure control remains unclear. To compare seizure control after surgical resection or radiosurgery for AVMs. We analyzed retrospectively collected information for 378 patients with cerebral AVMs treated at our institution from 1990 to 2010. The application of strict inclusion criteria resulted in a study population of 164 patients. In our cohort, 31 patients (20.7%) had Spetzler-Martin grade I AVMs, 51 (34.0%) grade II, 47 (31.3%) grade III, 20 (13.3%) grade IV, and 1 (0.7%) grade V. Of the 49 patients (30%) presenting with seizures, 60.4% experienced seizure persistence after treatment. For these patients, radiosurgery was associated with seizure recurrence (odds ratio: 4.32, 95% confidence interval: 1.24-15.02, P = .021). AVM obliteration was predictive of seizure freedom at last follow-up (P = .002). In contrast, for patients presenting without seizures, 18.4% experienced de novo seizures after treatment, for which surgical resection was identified as an independent risk factor (hazard ratio: 8.65, 95% confidence interval: 3.05-24.5, P < .001). Although our data suggest that achieving seizure freedom should not be the primary goal of AVM treatment, surgical resection may result in improved seizure control compared with radiosurgery for patients who present with seizures. Conversely, in patients without presenting seizures, surgical resection increases the risk of new-onset seizures compared with radiosurgery, but primarily within the early posttreatment period. Surgical resection and radiosurgery result in divergent seizure control rates depending on seizure presentation.


Metabolic adaptation for low energy throughput in orangutans.
Energy is the fundamental currency of life--needed for growth, repair, and reproduction--but little is known about the metabolic physiology and evolved energy use strategies of the great apes, our closest evolutionary relatives. Here we report daily energy use in free-living orangutans (Pongo spp.) and test whether observed differences in energy expenditure among orangutans, humans, and other mammals reflect known differences in life history. Using the doubly labeled water method, we measured daily energy expenditure (kCal/d) in orangutans living in a large indoor/outdoor habitat at the Great Ape Trust. Despite activity levels similar to orangutans in the wild, Great Ape Trust orangutans used less energy, relative to body mass, than nearly any eutherian mammal ever measured, including sedentary humans. Such an extremely low rate of energy use has not been observed previously in primates, but is consistent with the slow growth and low rate of reproduction in orangutans, and may be an evolutionary response to severe food shortages in their native Southeast Asian rainforests. These results hold important implications for the management of orangutan populations in captivity and in the wild, and underscore the flexibility and interdependence of physiological, behavioral, and life history strategies in the evolution of apes and humans.


The impact of lifestyle intervention on sedentary time in individuals at high risk of diabetes.
The Diabetes Prevention Program (DPP) lifestyle intervention successfully achieved its goal of increasing leisure physical activity levels. This current study examines whether the lifestyle intervention also changed time spent being sedentary and the impact of sedentary time on diabetes development in this cohort. 3,232 DPP participants provided baseline data. Sedentary behaviour was assessed via an interviewer-administered questionnaire and reported as time spent watching television specifically (or combined with sitting at work). Mean change in sedentary time was examined using repeated measures ANCOVA. The relationship between sedentary time and diabetes incidence was determined using Cox proportional hazards models. During the DPP follow-up (mean: 3.2 years), sedentary time declined more in the lifestyle than the metformin or placebo participants (p < 0.05). For the lifestyle group, the decrease in reported mean television watching time (22 [95% CI 26, 17] min/day) was greater than in the metformin or placebo groups (p < 0.001). Combining all participants together, there was a significantly increased risk of developing diabetes with increased television watching (3.4% per hour spent watching television), after controlling for age, sex, treatment arm and leisure physical activity (p < 0.01), which was attenuated when time-dependent weight was added to the model. In the DPP, the lifestyle intervention was effective at reducing sedentary time, which was not a primary goal. In addition, in all treatment arms, individuals with lower levels of sedentary time had a lower risk of developing diabetes. Future lifestyle intervention programmes should emphasise reducing television watching and other sedentary behaviours in addition to increasing physical activity. ClinicalTrials.gov NCT00004992.


Molecular dynamics study on the role of solvation water in the adsorption of hyperactive AFP to the ice surface.
Using computer simulations, the early stages of the adsorption of the CfAFP molecule to the ice surface were analyzed. We found that the ice and the protein interact at least as early as when the protein is about 1 nm away from the ice surface. These interactions are mediated by interfacial solvation water and are possible thanks to the structural ordering of the solvent. This ordering leads to positional preference of the protein relative to the ice crystal before the final attachment to the ice surface takes place, accompanied by the solidification of the interfacial water. It is possible because the solvation water of the ice-binding plane of CfAFP is susceptible to the overlapping with the solvation water of ice and is mostly changeable into ice itself. These remote interactions significantly increase efficacy of the adsorption process by facilitating the geometric adjustment of the active region of the CfAFP molecule to the ice surface. Because of the ordered nature of the water molecules at the ice-binding plane, the energy of their interactions with the ice-binding surface of the protein does not change upon the ongoing solidification of solvation water. However, the structure of the solvation water is not strictly ice-like and the growth of ice in the interfacial water is not initiated at the side of the protein. On the contrary, we find that solvation water of CfAFP solidifies slower than solvation water of ice - the solidification of interfacial water starts at the surface of ice. Moreover, in the presence of the CfAFP molecule, also solvation water of ice solidifies slower compared to the situation when the protein is not present next to the ice surface. Additionally, the presence of the protein molecule shifts the ratio of cubic to hexagonal ice that spontaneously forms at the ice surface, by introducing another layer of ordered water molecules - opposite to the ice lattice, at the other side of the crystallizing layer of water.


CE: Original Research: Helping Health Care Providers and Staff Process Grief Through a Hospital-Based Program.
: Background: Grief and loss are significant issues for health care providers, who may witness their patients' pain and suffering, experience feelings of grief as a result of caring for sick and dying people, and reexperience their own past losses. Unaddressed grief can lead to many issues for providers, their patients, and the health care system as a whole. Healing Loss: A Residential Workshop for Montefiore Associates is an experiential and educational program aimed at helping professional caregivers to identify and process grief and loss. Through retrospective analysis of program participation and feedback data, this study investigated the feasibility and effectiveness of offering an intensive bereavement support program to hospital employees in a large academic health system. Between 2013 and 2017, Montefiore Health System held nine Healing Loss workshops, serving 198 employees from diverse professions. These participants described multiple benefits, including being able to grieve more effectively, accessing support, and learning new tools for healing and self-care. Participants described the workshop experience as unique, cathartic, and life changing. The sustainability of the Healing Loss initiative during the four years of the study, together with strong feedback from participants, indicates that bereavement support for hospital employees is both feasible and beneficial.


Mechanism of polymerase II transcription repression by the histone variant macroH2A.
macroH2A (mH2A) is an unusual histone variant consisting of a histone H2A-like domain fused to a large nonhistone region. In this work, we show that histone mH2A represses p300- and Gal4-VP16-dependent polymerase II transcription, and we have dissected the mechanism by which this repression is realized. The repressive effect of mH2A is observed at the level of initiation but not at elongation of transcription, and mH2A interferes with p300-dependent histone acetylation. The nonhistone region of mH2A is responsible for both the repression of initiation of transcription and the inhibition of histone acetylation. In addition, the presence of this domain of mH2A within the nucleosome is able to block nucleosome remodeling and sliding of the histone octamer to neighboring DNA segments by the remodelers SWI/SNF and ACF. These data unambiguously identify mH2A as a strong transcriptional repressor and show that the repressive effect of mH2A is realized on at least two different transcription activation chromatin-dependent pathways: histone acetylation and nucleosome remodeling.


Lateral root morphogenesis is dependent on the mechanical properties of the overlaying tissues.
In Arabidopsis, lateral root primordia (LRPs) originate from pericycle cells located deep within the parental root and have to emerge through endodermal, cortical, and epidermal tissues. These overlaying tissues place biomechanical constraints on the LRPs that are likely to impact their morphogenesis. This study probes the interplay between the patterns of cell division, organ shape, and overlaying tissues on LRP morphogenesis by exploiting recent advances in live plant cell imaging and image analysis. Our 3D/4D image analysis revealed that early stage LRPs exhibit tangential divisions that create a ring of cells corralling a population of rapidly dividing cells at its center. The patterns of division in the latter population of cells during LRP morphogenesis are not stereotypical. In contrast, statistical analysis demonstrated that the shape of new LRPs is highly conserved. We tested the relative importance of cell division pattern versus overlaying tissues on LRP morphogenesis using mutant and transgenic approaches. The double mutant aurora1 (aur1) aur2 disrupts the pattern of LRP cell divisions and impacts its growth dynamics, yet the new organ's dome shape remains normal. In contrast, manipulating the properties of overlaying tissues disrupted LRP morphogenesis. We conclude that the interaction with overlaying tissues, rather than the precise pattern of divisions, is most important for LRP morphogenesis and optimizes the process of lateral root emergence.